Impaired wound healing after injury, surgery, or during disease impacts millions of patients each year, and better treatment strategies to enhance tissue repair are needed. In the skin, repair following injury involves a cascade of events that includes clotting, inflammation, wound closure via cell proliferation and migration, and re-vascularization and remodeling. Commonly, the process culminates with scarring, and regeneration of normal skin fails to occur. However, a recently popularized model for wound-induced hair neogenesis (WIHN), whereby new hair follicles and sebaceous glands regenerate in large acute and burn wounds, provides platform for studying how to modulate wound healing to promote more complete skin regeneration. Macrophages and fibroblasts are essential regulators of this process, and are involved in inflammation, wound closure, and regeneration of native skin structures. While soluble, biochemical factors such as chemokines, cytokines and growth factors in the wound environment are thought to regulate their responses, less is known about how biophysical cues regulate their function, despite the fact that cells exist within solid tissues that are rich in mechanical cues. Work from our laboratory and others have demonstrated that soft extracellular matrix hydrogels reduces macrophage inflammation and myofibroblast activation. Furthermore, we found that the a mechanically-activated and calcium permeable ion channel, plays a major role in mechanotransduction in both macrophages and fibroblasts. In this study, we propose to investigate role of stiffness and Piezo1 in skin wound healing. In Aim 1, we will investigate the roles of stiffness and Piezo1 on wound healing in murine full thickness skin wound. In Aim 2, we will examine the roles of stiffness and Piezo1 in regeneration using the WIHN model. An improved fundamental understanding of how cells sense their mechanical environment during wound healing may lead to new strategies that speed healing and enhance the quality of repaired tissue.